Lung growth factors in the amniotic fluid of normal pregnancies and with congenital diaphragmatic hernia.

AIM: Respiratory failure secondary to pulmonary hypoplasia is the main cause of death in congenital diaphragmatic hernia (CDH). Lung growth is regulated by growth factors (GFs), whose imbalances are reported in pathological conditions. We measured amniotic fluid levels of GFs, regulating lung development, in pregnancies with CDH and compared them with normal gestations. METHODS: Amniotic fluid was collected at amniocentesis and delivery from 4 women carrying fetuses with CDH and 12 with normal pregnancy. GFs were isolated and quantified. Same GFs were measured in lung biopsies collected during autopsy of three newborns dead of CDH. RESULTS: Impairment expression of lung GFs in the amniotic fluid of CDH pregnancies in comparison with normal was found. Fibroblast growth factor 10 (FGF10), fibroblast growth factor 7, vascular endothelial growth factor and transforming growth factor ß (TGFß) were decreased at amniocentesis, while platelet-derived growth factor (PDGF) increased. While FGF10 and PDGF tended to normalize at delivery, epidermal growth factor increased and TGFß was still decreased. Same GFs were similarly expressed in both lungs of babies dead of CDH. CONCLUSION: Anomalies in lung GFs expression of embryos and fetuses with CDH can be detected by measuring their levels in the amniotic fluid during pregnancy. Further investigation would help to correlate prenatal expression of GFs and clinical outcome of babies with CDH after birth.

Action of methotrexate and tofacitinib on directly stimulated and bystander-activated lymphocytes.

Chronic inflammation associated with autoimmune activation is characteristic of rheumatic diseases from childhood to adulthood. In recent decades, significant improvements in the treatment of these types of disease have been achieved using disease modifying anti-rheumatic drugs (DMARDs), such as methotrexate (MTX) and, more recently, using biologic inhibitors. The recent introduction of kinase inhibitors (for example, tofacitinib; Tofa) further increases the available ARDs. However, there are patients that do not respond to any treatment strategies, for whom combination therapies are proposed. The data regarding the combined action of different drugs is lacking and the knowledge of the mechanisms of ARDs and their actions upon pathogenic lymphocytes, which are hypothesized to sustain disease, is poor. An in vitro model of inflammation was developed in the current study, in which stimulated and unstimulated lymphocytes were cultured together, but tracked separately, to investigate the action of MTX and Tofa on the two populations. By analysing lymphocyte proliferation and activation, and cytokine secretion in the culture supernatants, it was established that, due to the presence of activated cells, unstimulated cells underwent a bystander activation that was modulated by the ARDs. Additionally, varying administration schedules were demonstrated to affect lymphocytes differently in vitro, either directly or via bystander activation. Furthermore, MTX and Tofa exerted different effects; while MTX showed an antiproliferative effect, Tofa marginally effected activation, although only a slight antiproliferative action, which could be potentiated by sequential treatment with MTX. Thus, it was hypothesized that these differences may be exploited in sequential therapeutic strategies, to maximize the anti­rheumatic effect. These findings are notable and must be accounted for, as bystander­activated cells in vivo could contribute to the spread of autoimmune activation and disease progression.